The 500 Days Of Summer star took to the stage on Thursday (January 26) to sing the Beatles track.
Gordon-Levitt previously collaborated with Zooey Deschanel on a cover of Nancy Wilson's 'What Are You Doing New Year's Eve?'
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He also teamed with The Dark Knight Rises co-star Anne Hathaway at his hitRECord annual Fall Formal in Los Angeles back in October.
The actor has a number of films due out this year, including Christopher Nolan's The Dark Knight Rises, Steven Spielberg's Lincoln and Quentin Tarantino's Django Unchained.
He also plays a killer named Joe in Looper, in which he'll wear prosthetics to resemble a young Bruce Willis.
Watch Joseph Gordon-Levitt singing 'Hey Jude' below:
